Summary
The replication control mechanism of Bacillus plasmid pRBH1 was analysed; pRBH1 contains four promoters, P1 to P4, and a large inverted repeat (63 base pairs) upstream of the protein (RepB) coding sequence. The stem and loop structure is surrounded by two promoters, P1 and P3, with different directions of transcription. One base substitution in the loop structure caused a change in copy number. Since the P1 promoter is located upstream of the replication origin of pRBH1, the transcript from the P1 promoter might serve as the primer of DNA replication. In vivo transcription from the P1 promoter was repressed by a trans-acting plasmid gene product. Since the RepB protein is involved in copy number control and RepB contains the consensus amino acid sequence of DNA binding proteins, RepB was thought to be the repressor. It was concluded from these data that the inverted repeat is involved in the control of copy number of the plasmid pRBH1. The RepB protein also contains two regions highly homologous with the Rom protein encoded on Escherichia coli plasmid ColE1. The possible mechanism for the copy number control of the plasmid via RepB protein and/or RNAs is discussed.
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References
Brückner R, Zyprian E, Matzura H (1984) Expression of a chloramphenicol resistance determinant carried on hybrid plasmids in gram-positive and gram-negative bacteria. Gene 32:151–160
Cesareni G, Muesing MA, Polisky B (1982) Control of ColE1 DNA replication: The rop gene product negatively affects transcription from the replication primer promoter. Proc Natl Acad Sci USA 79:6313–6317
Imanaka T, Fujii M, Aiba S (1981a) Isolation and characterization of antibiotic resistance plasmids from thermophilic bacilli and construction of deletion plasmids. J Bacteriol 146:1091–1097
Imanaka T, Tanaka T, Tsunekawa H, Aiba S (1981b) Cloning of the genes for penicillinase, penP and penI, of Bacillus licheniformis in some vector plasmids and their expression in Escherichia coli, Bacillus subtilis, and Bacillus licheniformis. J Bacteriol 147:776–786
Imanaka T, Fujii M, Aramori I, Aiba S (1982) Transformation of Bacillus stearothermophilus with plasmid DNA and characterization of shuttle vector plasmids between Bacillus stearothermophilus and Bacillus subtilis. J Bacteriol 149:824–830
Imanaka T, Ano T, Fujii M, Aiba S (1984) Two replication determinants of an antibiotic-resistance plasmid, pTB19, from a thermophilic bacillus. J Gen Microbiol 130:1399–1408
Imanaka T, Himeno T, Aiba S (1985) Effect of in vitro DNA rearrangement in the NH2-terminal region of the penicillinase gene from Bacillus licheniformis on the mode of expression in Bacillus subtilis. J Gen Microbiol 131:1753–1763
Light J, Molin S (1983) Post-transcriptional control of expression of the repA gene of plasmid R1 mediated by a small RNA molecule. EMBO J 2:93–98
Matsumura M, Katakura Y, Imanaka T, Aiba S (1984) Enzymatic and nucleotide sequence studies of a kanamycin-inactivating enzyme encoded by a plasmid from thermophilic bacilli in comparison with that encoded by plasmid pUB110. J Bacteriol 160:413–420
Mizuno T, Chou M-Y, Inouye M (1984) A unique mechanism regulating gene expression: Translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci USA 81:1966–1970
Muller RE, Ano T, Imanaka T, Aiba S (1986) Complete nucleotide sequences of Bacillus plasmids pUB110dB, pRBH1 and its copy mutants. Mol Gen Genet 202:169–171
Pabo CO, Sauer RT (1984) Protein-DNA recognition. Annu Rev Biochem 53:293–321
Scott JR (1984) Regulation of plasmid replication. Microbiol Rev 48:1–23
Som T, Tomizawa J (1983) Regulatory regions of ColE1 that are involved in determination of plasmid copy number. Proc Natl Acad Sci USA 80:3232–3236
Tomizawa J (1984) Control of ColE1 plasmid replication: The process of binding of RNA I to the primer transcript. Cell 38:861–870
Tomizawa J, Som T (1984) Control of ColE1 plasmid replication: Enhancement of binding of RNA I to the primer transcript by the Rom protein. Cell 38:871–878
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Communicated by P. Starlinger
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Ano, T., Imanaka, T. & Aiba, S. The copy number of Bacillus plasmid pRBH1 is negatively controlled by RepB protein. Molec Gen Genet 202, 416–420 (1986). https://doi.org/10.1007/BF00333271
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DOI: https://doi.org/10.1007/BF00333271